Total T-cell Research Articles

Cadmium-induced lung injury disrupts immune cell homeostasis in the secondary lymphoid organs in mice

Cadmium is a well-known toxic heavy metal that poses significant health risks, particularly through inhalation, smoking, and the consumption of contaminated food. Exposure to cadmium is linked to the development and exacerbation of chronic lung diseases such as pulmonary fibrosis and chronic obstructive pulmonary disease (COPD). This study investigated the systemic effects of intratracheal cadmium chloride (0.5mg/kg) instillation in C57BL/6 mice. All parameters, including inflammation assessment, lung function evaluation (using Flexi-vent), and immunophenotyping of T-cells in secondary lymphoid organs (spleen and mediastinal lymph nodes), were analyzed 14 days after Cd exposure. The results demonstrated that cadmium exposure led to significant immune cell infiltration in bronchoalveolar lavage (BAL) fluid, altered pro-inflammatory cytokine levels, and was associated with impaired lung function, characterized by increased lung resistance and Newtonian resistance. Analysis of T-cell populations revealed no significant changes in total T-cells in mediastinal lymph nodes and spleen, but a decrease in CD4+ T-cells and an increase in CD8+ T-cells were observed. These findings suggest that cadmium disrupts T-cell homeostasis in secondary lymphoid organs. Further research is crucial to elucidate the mechanisms underlying cadmium-induced lung injury and immune dysregulation, essential for developing effective therapeutic interventions against chronic lung diseases caused by cadmium exposure.

Unveiling the hidden link: elevated platelets and T cell subsets in 5% of moderate COVID-19 patients 48 days post-onset.

Platelets are hyperactived during acute COVID-19, promoting clotting and modulating immune-cell responses. Immune thrombocytopenia in adults can manifest as an uncommon complication resulting from various viral infections or as a rare adverse event associated with vaccination. However, their role in convalescent COVID-19 patients remains underexplored. This study examines platelet dynamics early in the pandemic, 48 days post-symptom onset, in unvaccinated patients. This longitudinal study included 298 unvaccinated COVID-19 patients (17 mild, 281 moderate) from multiple centers. Clinical evaluations and peripheral lymphocyte subset analyses via flow cytometry were conducted upon admission and on day 48 post-symptom onset (DPSO 48). At DPSO 48, 5.3% of moderate COVID-19 patients exhibited high platelet counts (>300×109/L), associated with elevated total T-cells (26.4%), CD4 T-cells (24.4%), CD8 T-cells (36.9%), and Tregs (33.9%) compared to patients with normal platelet counts. However, the CD4/CD8 T-cell ratio and T-cell subset frequencies remained unaffected, indicating ongoing T-cell homeostasis restoration. Additionally, a significant positive correlation (r=0.636, p=0.03) was found between platelet counts and B cells in patients with elevated platelet counts. Platelets may play a pivotal role in immune regulation during the recovery phase of COVID-19. Targeting platelets and their secreted mediators could improve immune balance in patients with immune disorders, highlighting a potential therapeutic approach for enhancing recovery in post-COVID-19 patients.

Characterisation of the tumour microenvironment in primary and recurrent glioblastomas.

Glioblastoma patients have a dismal prognosis, due to inevitable tumour recurrence and respond poorly to immunotherapy. Tumour-associated microglia/macrophages (TAMs) dominate the glioblastoma tumour microenvironment and have been implicated in tumour progression and immune evasion. Early recurrent glioblastomas contain focal reactive regions with occasional fibrosis, chronic inflammation, TAMs and tumour cells. Surgical specimens from these tumours are rare and provide crucial insights into glioblastoma recurrence biology. This study aimed to characterise TAM- and lymphocyte phenotypes in primary vs early- and late-recurrent glioblastomas. Patient-matched primary and recurrent glioblastomas were compared between patients with early recurrences (n = 11, recurrence ≤6months) and late recurrences (n = 12, recurrence after 12-19 months). Double-immunofluorescence stains combining Iba1 with HLA-DR, CD14, CD68, CD74, CD86, CD163, CD204 and CD206 along with stains for CD20, CD3, CD8 and FOXP3 were quantified with software-based classifiers. Reactive regions in early recurrent tumours contained more TAMs (31.4% vs 21.7%, P = 0.01), which showed increased expression of CD86 (59.4% vs 38.4%, P = 0.04), CD204 (48.5% vs 28.4%, P = 0.03), CD206 (25.5% vs 14.4%, P = 0.04) and increased staining intensity for CD163 (86.4 vs 57.7 arbitrary units, P = 0.02), compared to late recurring tumours. Reactive regions contained more B-lymphocytes compared to patient-matched primary tumours (0.71% vs 0.40%, P = 0.04). Fractions of total, cytotoxic and regulatory T-lymphocytes did not differ. Early recurrent glioblastomas showed enrichment for TAMs, expressing both pro- and anti-inflammatory markers and B-lymphocytes. This may indicate a time-dependent response to immunotherapy explained by time-dependent alterations in the immune-microenvironment in recurrent glioblastomas.

TUMOR INFECTION WITH HUMAN HERPES VIRUSES AND FEATURES OF PERIPHERAL BLOOD LYMPHOCYTE SUBSET COMPOSITION IN PATIENTS WITH UVEAL MELANOMA

Abstract Introduction. Currently, a role for human herpes viruses (HHV) in development of ocular oncopathology remains one of the poorly explored issues. An important factor contributing to neoplastic progression is impaired immunosurveillance particularly alterations in quantitative and qualitative composition of the hallmark peripheral blood lymphocyte subsets. Purpose: to determine and analyze peripheral blood lymphocyte subset composition in patients with uveal melanoma (UM) assessing human herpes viruses (HHV) infection in tumor material. Materials and Methods. Biomaterial from 99 patients with uveal tract tumors was examined by real-time polymerase chain reaction (rt-PCR) for DNA presence coupled to herpes virus type 1 and 2 (HSV-1,2), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), human herpes virus types 6 and 8 (HHV-6, HHV-8). A total of 231 test samples (tumor tissue (n=99), blood (n=132)) were examined. Commercial test-systems of Vector-Best CJSC were used for rt-PCR staging. Peripheral blood lymphocyte subset composition in all patients was studied by laser cytofluorometry on BD FACSCanto II flow cytometer. The control group included 33 healthy donors. Statistical data processing was performed using “Biostatd”, “Excel” (t - Student's t-test, level of statistical significance: p 0.05) software. Results. HHV DNA was detected in the tumor material from 11.3% of patients (n=11): in 72.7% of cases EBV, in 18.2% –– HHV-6. Co-infection with EBV and HHV-6 was detected in 1 case (retinal pigment epithelium adenocarcinoma). According to the PCR data, patients were divided into 2 groups based on tumor infectivity: Group 1 –– HHV “+” and Group 2 –– HHV “-”. Patients from HHV “+” group had significantly increased and decreased total T-cell (CD3+) and NK-cell count, respectively, compared with patients lacking HHV DNA in tumor tissue and control group. Individual analysis of frequencies deviating from normal range showed that HHV “+” group had 3-fold more often increased percentage of CD3+ lymphocytes, 2-fold more often absolute count of CD3+CD4+CD8+ cells, 2.3-fold more often rise in CD4+/CD8+ ratio. Conclusion. The obtained data suggest that viruses may be involved in maintaining immunological resistance in tumor patients.

Features of T-cell and humoral immunity of patients with acute coronary syndrome, with and without COVID-19, depending on/peripheral blood B-lymphocytes with the phenotype CD3–CD19+CD5+ level

The aim of the work was to assess the T-cell and humoral immune arms in patients with acute coronary syndrome (ACS) recovered from or not exposed to COVID-19. 86 men with ACS aged 40 to 65 years old, who suffered or not exposed to COVID-19, which required stenting of coronary arteries, were examined. Depending on the peripheral blood B-lymphocyte CD3–CD19+CD5+ level and the former COVID-19 history, all patients were divided into 6 groups. Of the previously COVID-19 exposed subjects CD3–CD19+CD5+ lymphocytes were at: reduced (group 1), normal (group 2), increased (group 3) level. COVID-19 unexposed subjects had CD3–CD19+CD5+ lymphocyte level: reduced (group 4), normal (group 5), increased (group 6). The most severe clinical manifestations were observed in group 1: with a larger rate of stent thrombosis and increased mortality. In absolute numbers, T-lymphocytes were significantly lower in group 1 compared to other groups, except group 4. The lowest both relative and absolute T-helper cell counts were recorded in covid-19 patients, with reduced CD3–CD19+CD5+ cell level. NK-lymphocytes both in relative and absolute level were significantly (p 0.05) higher in patients who suffered from COVID-19 as compared to those in patients unexposed to previous COVID-19. Percentage of late-activation T-regulatory cells was minimal in patients with former COVID-19 along with high B-lymphocyte CD3–CD19+CD5+ level, which significantly (p 0.05) differed from those in subjects lacking former COVID-19 history. In addition, significantly lower B-lymphocyte CD3–CD19+CD5– level was found in group 1 and 4. The IgM level peaked in group 5 — subjects lacking former COVID-19 history and having normal CD3–CD19+CD5+ cell level that significantly differed in comparison with that of in group 1, 2, 3. At the same time, the maximum value of IgG level was recorded in COVID-19 patients with normal CD3–CD19+CD5+ cells, and significantly differed from those found in group 5 and 6. C1-inhibitor level was higher in group 3 that significantly differed from that in other groups. SARS-CoV-2-specific IgG and IgM levels were significantly higher in COVID-19-positive vs. COVID-19-negative patients. Conclusion. In patients who have suffered COVID-19 with low B-lymphocyte CD3–CD19+CD5+ level, a significantly compromised immune protection was noted in comparison with other groups: a decline in total T-lymphocyte, T-helper, early- and late-activation T-lymphocyte, T-regulatory cell, B-lymphocyte CD3–CD19+CD5– levels, which was associated with a more severe disease course characterized by stent thrombosis and large mortality.

The effect of exercise and disease status on mobilization of anti-tumorigenic and pro-tumorigenic immune cells in women with breast cancer.

Mobilization of certain immune cells may improve the ability of the immune system to combat tumor cells, but the effect of acute exercise on mobilizing immune cells has been sparsely investigated in cancer patients. Therefore, we examined how acute exercise influences circulating immune cells in breast cancer patients. Nineteen newly diagnosed breast cancer patients aged 36-68 performed 30 minutes of moderate-intensity exercise with a cycle ergometer. Blood samples were collected at various time points: at rest, at 15 (E15) and 30 minutes (E30) after onset of the exercise, and at 30 and 60 minutes post-exercise. We analyzed several immune cell subsets using flow cytometry. Acute exercise increased the number of total leukocytes, neutrophils, lymphocytes, monocytes, basophils, total T-cells, CD4+ T-cells, T helper (Th) 2-cells, Th 17-cells, CD8+ T-cells, CD4-CD8- T-cells, CD56+ natural killer (NK) cells, and CD14-CD16+ monocytes. Many of the changes were transient. Proportions of NK-cells and CD8+ T-cells increased, while the proportion of myeloid derived suppressor cells (MDSCs) reduced, and proportion of regulatory T-cells remained unchanged by exercise. Several associations were detected between cell mobilizations and disease state. For instance, tumor size correlated negatively with NK cell mobilization at E15, and progesterone receptor positivity correlated negatively with CD8+ T-cell mobilization. The findings show that the proportions of CD8+ T-cells and NK cells increased and the proportion of MDSCs proportion decreased in breast cancer patients after 30-minute exercise, suggesting a change in the profile of circulating immune cells towards more cytotoxic/anti-tumorigenic. The mobilization of some immune cells also appears to be related to the disease state.

Adaptive post-COVID-19 immune response in female subjects of the Russian arctic region.

The Arctic region's unfavorable living conditions adversely affect the spread of infectious diseases, including COVID-19, This, in turn, can also lead to increased morbidity and mortality rates in the area due to anumber of factors such as climate, environment, and high prevalence rate of pre-existing health issues like diabetes, obesity, and respiratory infections. These circumstances adversely affect maintaining the level of working capability. The aim of this paper is to investigate the ratio of immunocompetent cells involved in the adaptive post-COVID-19 immune response. The research includes an immunological assessment of 29women aged 20-40 years residing in Arkhangelsk, Russia, six months after recovering from COVID-19. The count of leukocytes in the peripheral blood and their differential were evaluated using standard methods to assess the immunological status. To delve deeper into the immunological landscape, phenotypes of lymphocytes (CD5+, CD8+, CD10+, and CD95+) were evaluated using an indirect immunoperoxidase reaction with monoclonal antibodies on dried drop lymphocyte preparations. After incubating blood with latex molecules, the activity and quantity of phagocytes were assessed using alight microscope. The neutrophil/lymphocyte ratio was found to be inverted in the female subjects under investigation. The high concentration of cytotoxic T-lymphocytes (CD8+) and lymphocytes with apoptotic receptors (CD95+) suggests apotential correlation with aconcurrent reduction in the expression of the total T-cell marker (CD5+) across all cases. This association was further linked to adecrease in lymphoproliferative activity and arelative decline in phagocytic activity. These findings led us to posit that the total recovery time after COVID-19 might extend beyond six months, indicative of aprolonged impact on the body's protective capacity. Our observations prompt the hypothesis that cellular immunity plays acrucial role in determining the severity of COVID-19 infection. Specifically, individuals with initially robust phagocytic activity may be predisposed to experiencing amilder form of the infection. However, this assumption warrants further investigation and clarification in individuals with moderate and severe disease progression (Tab. 1, Ref. 17). Keywords: arctic, COVID-19, cytotoxic t-lymphocytes, apoptosis, lymphoproliferation, cellular immunity, phagocytic activity.

Dynamics of pulmonary mucosal cytotoxic CD8 T-cells in people living with HIV under suppressive antiretroviral therapy

BackgroundDespite the success of antiretroviral therapy (ART), people living with HIV (PLWH) suffer from a high burden of pulmonary diseases, even after accounting for their smoking status. Cytotoxic CD8 T-cells are likely implicated in this phenomenon and may act as a double-edged sword. While being essential in viral infection control, their hyperactivation can also contribute to lung mucosal tissue damage. The effects of HIV and smoking on pulmonary mucosal CD8 T-cell dynamics has been a neglected area of research, which we address herein.MethodsBronchoalveolar lavage (BAL) fluid were obtained from ART-treated PLWH (median duration of supressed viral load: 9 years; smokers: n = 14; non-smokers: n = 21) and HIV-uninfected controls (smokers: n = 11; non-smokers: n = 20) without any respiratory symptoms or active infection. Lymphocytes were isolated and CD8 T-cell subsets and homing markers were characterized by multiparametric flow cytometry.ResultsBoth smoking and HIV infection were independently associated with a significant increase in frequencies of total pulmonary mucosal CD8 T-cell. BAL CD8 T-cells were primarily CD69 + expressing CD103 and/or CD49a, at least one of the two granzymes (GzmA/GzmB), and little Perforin. Higher expression levels of CD103, CD69, and GzmB were observed in smokers versus non-smokers. The ex vivo phenotype of GzmA + and GzmB + cells revealed increased expression of CD103 and CXCR6 in smokers, while PLWH displayed elevated levels of CX3CR1 compared to controls.ConclusionSmoking and HIV could promote cytotoxic CD8 T-cell retention in small airways through different mechanisms. Smoking likely increases recruitment and retention of GzmB + CD8 Trm via CXCR6 and CD103. Heightened CX3CR1 expression could be associated with CD8 non-Trm recruitment from the periphery in PLWH.

Spatially mapping the single-cell immune landscapes of melanoma brain metastases using imaging mass cytometry.

e21517 Background: Up to 60% of metastatic melanoma patients develop brain metastases, and 5% develop leptomeningeal disease (LD), which presents with the worst prognosis of any melanoma brain metastasis (MBM) infiltration pattern. Median survival duration for MBM is ~12 months, however for LD patients it can be < 10 weeks. The spatial immune landscape of MBM with and without LD remains poorly understood. Methods: To spatially characterize the tumor microenvironment (TME) of MBM, we performed CyTOF Imaging Mass Cytometry (CyTOF-IMC) on a highly multiplexed panel of 35 antibodies for 21 MBMs (13 with LD, 8 without LD). We performed a novel cell segmentation, cell type assignment and identification approach to identify cell lineages to spatially characterize the TME of MBMs, segmenting and classifying 130,000 cells into 19 cell types. Results: We found enrichment of dendritic cells in the direct neighbourhood of melanoma cells of patients with LD (p = 0.019) and trends towards elevation of anti-tumoral monocyte populations in the TME of patients without LD. To investigate survival outcomes, we divided our cohort into long (≥365days) and short-survivors ( < 365 days), and found significant elevation of anti-tumoral T-cell populations (Total T-cells, CD8+ T-cells, CD4+ T-cells and CD3+CD4-CD8- T-cells, p < 0.05), and closer proximity of T-cells (CD8+ & CD4+ T-cells) to melanoma cells (p < 0.05). Survival analysis using median cell-proportion for each T-cell subset showed statistically significant (p < 0.05) survival differences between patients with high and low cell-proportions of CD8+ T-cells and CD4+ T-cells, with similar trends observed for a subset of patients (n = 13) with LD and a subset of patients treated with ICIs (n = 10), and results were confirmed by a Cox Regression Model. Cellular neighborhood (CN) analysis was performed generating N = 12 stable CNs. We found patients with higher proportions of CN9 (M1-like-microglia/M2-like-microglia/melanoma), CN10 (M1-like-macrophages/M2-like-macrophages/melanoma) and CN12 (Cytotoxic-T-cells/Helper-T-cells/melanoma) was associated with longer overall survival after diagnosis with MBM (p < 0.01). Conclusions: Proportion and proximity of anti-tumoral monocyte and T-cell populations may play a key role in modulating tumor response and overall survival for patients with MBM, including for patients with LD or for patients treated with ICIs.

Effect of UNO101 on tumor microenvironment in relapsed or refractory, unresectable, primary, or metastatic cutaneous and subcutaneous malignancies.

e14575 Background: Nitric oxide (NO) is a ubiquitous, endogenously generated gas implicated in the homeostatic regulation of physiological processes, and NO is implicated in numerous pathological conditions. Preclinical studies evaluating the effect of high concentration exogenously administered NO demonstrated anti-cancer properties and suggested that it may serve as a potent tumoricidal agent. We have previously shown that treating mouse colon carcinoma (CT26) tumor-bearing mice with ultra-high concentrations of nitric oxide (UNO) upregulates innate and adaptive immune cells locally and systemically. Methods: Upon a signed written ICF, a single UNO101 dose of 25,000 parts per million (ppm), administered intratumorally over 5 minutes, was evaluated in relapsed or refractory, unresectable, primary, or metastatic cutaneous and subcutaneous malignancies. Subjects with an ECOG PS of 0–3, at least 3 months of life expectancy and tumor size of 4.5 mm minimum and 30 mm maximum length were eligible for enrollment. RECIST v1.1 and clinical physical examination by PI assessment was utilized to assess the rate of malignant tumor response. Toxicity was graded per NCI CTCAE v5.0. Results: In this first-in-human Phase 1 clinical trial, six subjects, 4F/2M were enrolled with a median age of 61.5 years, (range, 34–81) and ECOG PS reported as 0 or 1. Tumor types enrolled included breast (n=3), melanoma (n=1), and squamous cell carcinoma (n=2) with a median of 4 prior lines of therapy (range, 2–11) and a median of 4.7 years (range, 1.4–9.5) since time of diagnosis to study entry. One related SAE (not dose limiting) of hypoxia, Grade 4, was reported, which resulted in cessation of UNO101 treatment after 1:55 minutes. The other five administrations were completed successfully, with no further SAEs. Other reported drug related AEs were primarily Grade 1 that resolved/recovered without sequelae. Clinical immune blood biomarkers post UNO101 treatment demonstrated modulation of the tumor microenvironment by upregulating of cytotoxic T-cells (11.1%), T-central memory cells (190.6%) as a percent of total T-cells, and dendritic cells (231.4%) as a percentage of WBCs, from baseline to Day 21. Further, T-regulatory cells (38.0%) were downregulated as a percentage of total T-cells from baseline to Day 21. Based on available physical examination and caliper measurements, clinical presentation on Day 7 on two subjects, significant clinical responses were observed. Conclusions: To date,a single dose of UNO101 at 25,000 ppm is well tolerated, with biomarkers suggesting immune action within the tumor microenvironment, trending in a favorable direction on Day 21 in a heavily pre-treated population. Additional UNO101 single dose of 50,000 ppm, and possibly 100,000 ppm, will be further evaluated for safety/tolerability, impact on immune biomarkers, and preliminary efficacy. Clinical trial information: NCT05351502 .

A standardized combination of Terminalia chebula and Withania somnifera extracts enhances immune function in adults: a pilot randomized, double-blind, placebo-controlled clinical study.

The use of botanical medicine has been demonstrated as a potential strategy to manage or treat a variety of health issues. Terminalia chebula (Retz) fruit and Withania somnifera (L.) Dunal roots are important medicinal herbs described in Ayurveda and traditional therapy for diverse health benefits. This pilot study aimed to evaluate the immune function-enhancing potential of a unique blend of T. chebula fruit and W. somnifera root extracts, LN20189, in healthy men and women. Forty healthy volunteers (age: 35-60 years) were randomized into two groups receiving either LN20189 (500 mg per day) or a matched placebo over 28 consecutive days. The total T-cell population was the primary efficacy measure in this study. The secondary efficacy measures included counts of CD4, CD8, natural killer (NK) cells, serum levels of interleukin-2 (IL-2), interferon-gamma (IFN-γ), total immunoglobulin-G (IgG), and Immune Function Questionnaire (IFQ) scores. Safety parameter assessments were also conducted. Post-trial, in LN20189-supplemented subjects, T cells, CD4, NK cells count, and the CD4:CD8 ratio were increased by 9.32, 10.10, 19.91, and 17.43%, respectively, as compared to baseline. LN20189 supplementation increased serum IFN-γ and IgG levels by 14.57 and 27.09% from baseline and by 13.98 and 21.99%, compared to placebo, respectively. Also, the IFQ scores in the LN20189 group were 84.68% (vs. baseline) and 69.44% (vs. placebo) lower at the end of the trial. LN20189 improved the study volunteers' cellular and humoral immune functions. In summary, LN20189 supplementation was found tolerable and improved the key cellular and humoral factors of the immune system and helped improve immune function of the trial volunteers.

Decreased but persistent epigenetic age acceleration is associated with changes in T-cell subsets after initiation of highly active antiretroviral therapy in persons living with HIV.

Persons living with HIV (PLWH) experience the early onset of age-related illnesses, even in the setting of successful human immunodeficiency virus (HIV) suppression with highly active antiretroviral therapy (HAART). HIV infection is associated with accelerated epigenetic aging as measured using DNA methylation (DNAm)-based estimates of biological age and of telomere length (TL). DNAm levels (Infinium MethylationEPIC BeadChip) from peripheral blood mononuclear cells from 200 PLWH and 199 HIV-seronegative (SN) participants matched on chronologic age, hepatitis C virus, and time intervals were used to calculate epigenetic age acceleration, expressed as age-adjusted acceleration residuals from 4 epigenetic clocks [Horvath's pan-tissue age acceleration residual (AAR), extrinsic epigenetic age acceleration (EEAA), phenotypic epigenetic age acceleration (PEAA), and grim epigenetic age acceleration (GEAA)] plus age-adjusted DNAm-based TL (aaDNAmTL). Epigenetic age acceleration was compared for PLWH and SN participants at two visits: up to 1.5years prior and 2-3years after HAART (or equivalent visits). Flow cytometry was performed in PLWH and SN participants at both visits to evaluate T-cell subsets. Epigenetic age acceleration in PLWH decreased after the initiation of HAART but remained greater post-HAART than that in age-matched SN participants, with differences in medians of 6.6, 9.1, and 7.7 years for AAR, EEAA, and PEAA, respectively, and 0.39 units of aaDNAmTL shortening (all p < 0.001). Cumulative HIV viral load after HAART initiation was associated with some epigenetic acceleration (EEAA, PEAA, and aaDNAmTL), but even PLWH with undetectable HIV post-HAART showed persistent epigenetic age acceleration compared to SN participants (p < 0.001). AAR, EEAA, and aaDNAmTL showed significant associations with total, naïve, and senescent CD8 T-cell counts; the total CD4 T-cell counts were associated with AAR, EEAA, and PEAA (p = 0.04 to <0.001). In an epigenome-wide analysis using weighted gene co-methylation network analyses, 11 modules demonstrated significant DNAm differences pre- to post-HAART initiation. Of these, nine were previously identified as significantly different from pre- to post-HIV infection but in the opposite direction. In this large longitudinal study, we demonstrated that, although the magnitude of the difference decreases with HAART is associated with the cumulative viral load, PLWH are persistently epigenetically older than age-matched SN participants even after the successful initiation of HAART, and these changes are associated with changes in T-cell subsets.

Indicators of Immunological Status in Pregnant Women with Chronic Viral Hepatitis B and C

The work presents the results of a study of immunological indicators of cellular and humoral immunity in 45 pregnant women with chronic viral hepatitis B and C. Immunological parameters in pregnant women with CHB and CHC were characterized by suppression of the cellular component of immunity, which is manifested by a decrease in the number of total T-lymphocytes (CD3+), T-helpers (CD4+) and T-suppressors (CD8+), as well as natural killer cells (CD16), which is associated with the mechanism of maintaining pregnancy. Changes in humoral immunity are characterized by a moderate increase in the level of IgG and CEC; in case of CHC, relapse of the disease was also accompanied by an increase in class M immunoglobulins. Chronic viral hepatitis B and C in pregnant women leads to secondary immunodeficiency and an increase in the level of viral replication. A direct correlation has been revealed between indicators of the cellular immune response in CHB and CHC with the level of viral load, which must be taken into account when managing pregnancy.

NKX2‑1 copy number alterations are associated with oncogenic, immunological and prognostic remodeling in non‑small cell lung cancer.

NK2 homeobox 1 (NKX2-1) copy number alterations (CNAs) are frequently observed in lung cancer. However, little is known about the complete landscape of focal alterations in NKX2-1 copy number (CN), their clinical significance and their therapeutic implications in non-small cell lung cancer (NSCLC). The correlations between NKX2-1 expression and EGFR driver mutations and programmed death ligand 1 (PD-L1) co-expression were studied using immunohistochemistry and PCR from the tumors of recruited Filipino patients (n=45). Clinical features of NSCLC with NKX2-1 CNAs were resolved at the tumor and clonal levels using the molecular profiles of patients with lung adenocarcinoma and lung squamous cell carcinoma from The Cancer Genome Atlas (n=1,130), and deconvoluted single-cell RNA-seq data from the Bivona project (n=1,654), respectively. Despite a significant and positive correlation between expression and CN (r=0.264; P<0.001), NKX2-1 CNAs exerted a stronger influence on the combined EGFR and PD-L1 status of NSCLC tumors than expression. NKX2-1 CN gain was prognostic of favorable survival (P=0.018) and a better response to targeted therapy. NKX2-1 CN loss predicted a worse survival (P=0.041). Mutational architecture in the Y-chromosome differentiated the two prognostic groups. There were 19,941 synonymous mutations and 1,408 genome-wide CN perturbations associated with NKX2-1 CNAs. Tumors with NKX2-1 CN gain expressed lymphocyte markers more heterogeneously than those with CN loss. Higher expression of tumor-infiltrating lymphocyte gene signatures in CN gain was prognostic of longer disease-free survival (P=0.005). Tumors with NKX2-1 CN gain had higher B-cell (P<0.001) and total T-cell estimates (P=0.003). NKX2-1 CN loss was associated with immunologically colder tumors due to higher M2 macrophage infiltrates (P=0.011) and higher expression of immune checkpoint proteins, CD274 (P=0.025), VTCN1 (P<0.001) and LGALS9 (P=0.002). In conclusion, NKX2-1 CNAs are associated with tumors that exhibit clinically diverse characteristics, and with unique oncogenic, immunological and prognostic signatures.

Chimeric Antigen Receptor T-Cell Recruitment and Killing can be Evaluated on an Organ-Chip model system

Abstract Objective. Chimeric antigen (CAR) T-cell therapies for solid tumors are challenging to develop due to the lack of human relevant models that adequately capture the mechanisms of CAR T-cell recruitment-a critical yet often overlooked rate-limiting step. The current study aimed to develop a novel system for investigating both the recruitment and killing capacity of CAR T cells in Organ-Chips. Methods. Chip-S1™ Stretchable Chips (Emulate) were seeded with a human HER2+ non-small cell lung cancer cell line (NSCLC) in the top channel and lung microvascular endothelial cells in the vascular (bottom) channel to create a tumor-vascular interface. Both channels were primed with an NSCLC-specific mix of chemokines and cytokines. HER2-specific CAR T cells were perfused through the vascular channel. Total CAR T-cell recruitment into the top channel and CAR T-mediated killing were measured over 48 hours. The IL-2 cytokine was also co-administered with CAR T cells as immunotherapy. Results. HER2 CAR T cells demonstrated significant antigen-specific killing when recruited to the top channel as measured by caspase activity and cytokine release compared to the control. Furthermore, IL-2 co-therapy increased CAR T-cell recruitment and antigen-specific killing compared to the control group. Conclusion. These findings suggest that human-centric Organ-Chips can evaluate CAR T-cell therapies for solid tumors by integrating the critical rate-limiting steps of CAR T recruitment and killing.

Immune system parameters in patients with chronic opistorchiasis based on genetic polymorphism associated with carbohydrate and lipid metabolism disorders

A study assessing differences in the immunopathogenesis of chronic opisthorchiasis with or without genetic polymorphisms in patients associated with predisposition to type II diabetes mellitus and lipid metabolism disorders was carried out. Venous blood samples from 89 patients were collected to analyze immunological parameters and genetic polymorphisms by using pyrosequencing. In the presence of mutations affecting regulation of carbohydrate metabolism, patients with opisthorchiasis had less pronouncedly increased phagocytic activity, a lower monocyte count, higher total T-lymphocyte count, lower T-cytotoxic cell and B-lymphocyte counts, lower IgA, IgG but higher IgM concentration. This indicates a moderately compromised nonspecific resistance, imbalanced effector T- and B-immunity, but in most cases (PPARG, TCF7L2 rs12255372, CDKAL1, CDKN2A/2B, SLC30A8) aggravated humoral immune reaction to invasion particularly revealed by lower B-lymphocyte count. The presence of polymorphisms that alter lipid metabolism regulation bidirectionally affects the parameters of immune response. An increased B-lymphocyte count and other indicators of humoral immune activation to invasion, which are detected in groups with mutations in the APOE (rs429358), PCSK9, ABCA1, APOC3 rs2854117 genes, can contribute to a more effective response to sustained antigenic stimulation. Changes in the T-lymphocytes subpopulations, characteristic of opisthorchiasis invasion, are aggravated in the presence of mutations in the PCSK9, ABCA1, and APOC3 rs2854117 genes. Mutations in the genes APOC3 rs5128, LPL rs268 activate patients’ nonspecific resistance, although this effect may also be associated with exhaustion of neutrophil bactericidal reserve. In general, minor alleles of the APOE (rs429358), APOC3 rs2854117, LPL rs268, LPL rs328, PON1 rs662 genes can be considered “protective” for the immunopathogenesis of chronic opisthorchiasis. Thus, patients with chronic opisthorchiasis with different genotypes predisposing to carbohydrate and lipid metabolism disorders had significant differences in immune system parameters, which can also affect the disease course. Mutations in different loci of the PCSK9, ABCA1, APOE, APOC3, and PON1 genes have opposite effects on the analyzed immune parameters. In the presence of mutations in other genes (PPARG, TCF7L2 rs12255372, CDKAL1, CDKN2A/2B, SLC30A8, LPL), opisthorchiasis invasion leads to more pronounced alterations in immune response; mutations in the lipoprotein lipase gene may have some “protective” immune-related effect in opisthorchiasis. The effect of all studied genetic polymorphisms associated with a predisposition to developing type II diabetes mellitus was predominantly negative.

Correlation of immune fitness with response to teclistamab in relapsed/refractory multiple myeloma in the MajesTEC-1 study

AbstractTeclistamab, an off-the-shelf B-cell maturation antigen (BCMA) × CD3 bispecific antibody that mediates T-cell activation and subsequent lysis of BCMA-expressing myeloma cells, is approved for the treatment of patients with relapsed/refractory multiple myeloma (R/RMM). As a T-cell redirection therapy, clinical outcomes with teclistamab may be influenced by patient immune fitness and tumor antigen expression. We correlated tumor characteristics and baseline immune profiles with clinical response and disease burden in patients with R/RMM from the pivotal phase 1/2 MajesTEC-1 study, focusing on patients treated with 1.5 mg/kg of teclistamab (N = 165). Peripheral blood samples were collected at screening, and bone marrow samples were collected at screening and cycle 3. Better clinical outcomes to teclistamab correlated with higher baseline total T-cell counts in the periphery. In addition, responders (partial response or better) had a lower proportion of immunosuppressive regulatory T cells (Tregs), T cells expressing coinhibitory receptors (CD38, PD-1, and PD-1/TIM-3), and soluble BCMA and a T-cell profile suggestive of a more cytolytic potential, compared with nonresponders. Neither frequency of baseline bone marrow BCMA expression nor BCMA-receptor density was associated with clinical response to teclistamab. Improved progression-free survival was observed in patients with a lower frequency of T cells expressing exhaustion markers and immunosuppressive Tregs. Overall, response to teclistamab was associated with baseline immune fitness; nonresponders had immune profiles suggestive of immune suppression and T-cell dysfunction. These findings illustrate the importance of the contribution of the immune landscape to T-cell redirection therapy response. This trial was registered at www.ClinicalTrials.gov as #NCT03145181/NCT04557098.

Abstract CT208: High tumor monocyte content predicts long-term survival for patients with operable oesophageal cancer treated with neoadjuvant immunochemotherapy in the LUD2015-005 trial

Abstract Background: Combined immune checkpoint inhibitors (ICI) with chemotherapy in advanced oesophageal cancer (OC) has shown survival benefit in phase III studies, resulting in ICI’s recent approval to treat advanced OC. The role of ICI in peri-operative OC is debated, and large clinical studies are ongoing. Patients’ response to immunochemotherapy is highly variable and it is unclear what underlies this heterogeneity. To address this challenge, 38 advanced and 35 operable OC patients were enrolled to receive ICI in the LUD2015-005 trial (NCT02735239). Methods: We investigated the clinical and biological effects of adding the anti-PD-L1 ICI durvalumab to peri-operative chemotherapy or chemoradiotherapy (CRT) in a phase II multicentre study (LUD2015-005 trial). Operable OC patients received ICI for four weeks, followed by combination treatment with ICI + chemotherapy (oxaliplatin and capecitabine or fluorouracil, leucovorin, oxaliplatin, and docetaxel (FLOT)), or ICI + CRT (CROSS regimen). Patients could also receive ICI for up to 6 months after surgery. Tissue samples were collected at baseline, after four weeks ICI-only, and on the day of surgery. Further to assessing synergy of ICI with standard peri-operative treatment for OC, our trial design enables an in-depth analysis of molecular features of response, and to dissect the impact of ICI alone on the tumor microenvironment. Single cell and bulk transcriptomic data were generated and used to perform RNA deconvolution to estimate tumor cell composition. Results: The treatment was well-tolerated and two-year survival was 80% and 73.3% with ICI + chemotherapy and ICI + CRT, respectively. Survival was superior to that of propensity score matched patients receiving standard of care (p = 0.047). Tumor cell composition analysis revealed that neither total T-cell numbers nor a specific T-cell functional phenotype was associated with outcome, but high tumor monocyte content (TMC) at baseline was highly predictive of overall survival (p = 0.003). Moreover, a 13-gene core monocyte gene signature was sufficient to predict overall survival (p = 0.022) and major pathological response (mPR, Mandard 1-2, p = 0.009). Patients with mPR at resection showed enrichment of interferon and IL15 signalling at the four-week ICI-only timepoint (p &amp;lt; 0.001). Single cell trajectory interference combined with immune response enrichment analysis also show that high interferon and IL15 signalling drive monocytes to differentiate into M1 macrophages and dendritic cells. Conclusion: Similar to our previous finding in immunochemotherapy treated advanced OC patients (Carroll et al. Cancer Cell 2023), TMC is a highly predictive biomarker of long-term survival for operable OC patients treated with neoadjuvant immunochemotherapy, highlighting the importance of tumor monocytes and their differentiation potential in OC patients’ response to ICI. Citation Format: Hannah S. Fuchs, Sabrina A. James, Thomas M. Carroll, Phil F. Xie, Joseph A. Chadwick, Duncan Parkes, Simon R. Lord, Lucinda Griffiths, Tim Underwood, Ioannis Karydis, Russell T. Petty, Benjamin Schuster-Böckler, Richard P. Owen, Mark R. Middleton, Xin Lu. High tumor monocyte content predicts long-term survival for patients with operable oesophageal cancer treated with neoadjuvant immunochemotherapy in the LUD2015-005 trial [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT208.

Роль клеточного иммунитета в развитии злокачественных новообразований у лиц, подвергшихся хроническому облучению

Some long-term effects of radiation could be related to changes in the immune system resulting from radiation exposure. Immunity disorders caused by radiation can influence carcinogenesis. Cellular immunity factors were investigated in peripheral blood of workers chronically exposed to occupational combined radiation (external gamma-rays and internal alpha-particles), with malignant neoplasms diagnosed after blood samples were taken or without them, and in the control group. The aim of this study was examine effects of radiation on the cellular immunity status in individuals chronically exposed to ionizing radiation who had malignant neoplasms developed after blood sampling. The relative and absolute number of lymphocyte subpopulations (total T-cells, T-helpers, T-cytotoxic, total B-cells, NK-cells, NKT-cells and activated T-cells) was detected by flow cytofluorometry. The absolute number of T-cells was significantly reduced in workers chronically exposed to occupational combined irradiation, with or without malignancies, compared to the control, which may contribute to tumor progression at an early stage of its onset. At the same time, workers without malignancies had a significant increase in the relative number of T-cytotoxic lymphocytes, which may be a factor preventing tumor development. A significant increase in the relative number of natural killer cells (NK cells) was detected in individuals with malignant neoplasms chronically exposed to occupational combined irradiation, compared with the control, which may indicate enhanced antitumor defense that developed in response to exposure to tumor antigens. In addition, a significant decrease in the absolute and relative number of T- and B-lymphocytes was found in the group of workers with malignant neoplasm, compared with the control. A significant increase in the relative number of T-helpers was found in both groups of workers. Since the role of T-helpers in the antitumor response is ambiguous, additional research on types of T-helpers is planned to clarify the results of the present study

Role of cellular immunity in malignant tumors development in individuals chronically ex-posed to ionising radiation

Some long-term effects of radiation could be related to changes in the immune system resulting from radiation exposure. Immunity disorders caused by radiation can influence carcinogenesis. Cellular immunity factors were investigated in peripheral blood of workers chronically exposed to occupational combined radiation (external gamma-rays and internal alpha-particles), with malignant neoplasms diagnosed after blood samples were taken or without them, and in the control group. The aim of this study was examine effects of radiation on the cellular immunity status in individuals chronically exposed to ionizing radiation who had malignant neoplasms developed after blood sampling. The relative and absolute number of lymphocyte subpopulations (total T-cells, T-helpers, T-cytotoxic, total B-cells, NK-cells, NKT-cells and activated T-cells) was detected by flow cytofluorometry. The absolute number of T-cells was significantly reduced in workers chronically exposed to occupational combined irradiation, with or without malignancies, compared to the control, which may contribute to tumor progression at an early stage of its onset. At the same time, workers without malignancies had a significant increase in the relative number of T-cytotoxic lymphocytes, which may be a factor preventing tumor development. A significant increase in the relative number of natural killer cells (NK cells) was detected in individuals with malignant neoplasms chronically exposed to occupational combined irradiation, compared with the control, which may indicate enhanced antitumor defense that developed in response to exposure to tumor antigens. In addition, a significant decrease in the absolute and relative number of T- and B-lymphocytes was found in the group of workers with malignant neoplasm, compared with the control. A significant increase in the relative number of T-helpers was found in both groups of workers. Since the role of T-helpers in the antitumor response is ambiguous, additional research on types of T-helpers is planned to clarify the results of the present study.